Nano-liquid chromatography is a desirable technique because when coupled with mass spectrometry it can identify many of the components of a very complex proteomic or glycomic sample including subtle chemical variations within the sample, and can also obtain quantitative information. The technique is renowned for its high chromatographic resolution, high sensitivity, and small sample size requirement. But the technique remains cumbersome as it suffers from several limitations including system reproducibility issues created by user inabilities in column and/or spray emitter set-up, difficulty making nanofluidic connections, and difficulty troubleshooting system malfunctions. Others have tried to simplify this underutilized technique through primarily chip-based approaches, but have unfortunately compromised the technique's resolution and sensitivity. However, in a Phase I project, CorSolutions developed a robust, reliable, user-friendly nanoLC/ESI device that addresses these limitations of conventional nanoLC/ESI systems while still maintaining state-of- the-art performance, and is able to operate at elevated UHPLC pressures. These Phase I prototypes are the first plug-and-play nanoLC/ESI devices which do not compromise the high chromatographic resolution and sensitivity for which nanoLC is renowned. But although the technology developed in Phase I offers great improvements over conventional approaches, further developments are required for the technology to reach its large commercialization potential. The goals of the Phase II are to build upon the compression connection, plug-and-play technology developed in Phase I, in order to ready the technology for commercialization. This includes development of a cartridge-based insert for the column, allowing a column of any length to be wound in a small footprint. This enhancement will allow the overall device dimension to be substantially reduced. Additionally an emitter changer will be added, permitting the device to quickly exchange an emitter, in less than 1 sec, without need for user intervention. A spray sensor and leak sensors will be incorporated into the device with "smart" software to continuously monitor the sensor read back. The software will be capable of diagnosing system malfunctions and attempting automated repair such as an emitter change, greatly simplifying the troubleshooting process. Furthermore the ability to deliver nebulization gas will be incorporated into the device. These enhancements will allow for product commercialization. We see this simple, intuitive product becoming an integral part of biological laboratories, allowing standardization of proteomic and glycomic analyses. And most importantly the product will allow biologists to benefit from the highly informative nanoLC/MS data, enabling scientific breakthroughs and greater understanding of complex biological systems. This significant reduction in complexity of nanoLC will also allow for the technique to infiltrate deeper into the life sciences. PUBLIC HEALTH RELEVANCE: Nano-liquid chromatography mass spectrometry (nanoLC/MS) is a powerful technique for proteomic and glycomic studies which is able to not only identify proteins and glycans from very complex mixtures, but is also able to structurally characterize differences between the components, which is particularly important for post-translational modifications. However, conventional nanoLC/MS is a very difficult technique to perform, requiring a highly skilled user. Therefore this powerful technique, which could enable scientific discoveries and greater understanding of complex biological systems, is alarmingly underutilized. CorSolutions proposes to develop an intuitive, robust, reliable, user-friendly, "plug-and-play" device, which will address the challenges of conventional nanoLC/MS, dramatically reducing the complexity of the technique and thereby allowing its pervasive use in biological laboratories.